AGM STUDIES: A crucial element in understanding the pathogenesis of lentiviral infections is to compare and contrast parameters between progressive and non-progressive infections. Indeed, elucidating the mechanisms underlying AIDS resistance of SIV-infected natural hosts may provide critical information to better understand AIDS pathogenesis. To date, >40 primate lentiviruses have been found to naturally infect African nonhuman primate hosts at high prevalence levels. In general, SIVs infecting African species evolved through host-dependent evolution, meaning that they have infected different NHP species for thousands, or hundreds of thousands, of years, possibly since primate speciation. During this long history, SIVs and their natural hosts evolved to coexistence, with the African NHPs infected with their species-specific viruses generally not showing signs of SIV-induced disease or simian AIDS. To determine mechanisms by which natural host African nonhuman primates are able to survive SIV infection without progression to chronic infection we have studied the phenotypes, frequencies, functionality, and SIV-infection frequency within individual T cell subsets of SIVagm-infected and uninfected African green monkeys. We believe these studies will lead to an understanding of how these animals are able to live with SIVagm and not progress through chronic infection and avoid simian AIDS. We have found that (1) Most CD4 T cells from African green monkeys down-regulate CD4 in vivo as they enter the memory pool, (2) Down regulation of CD4 by memory T cells is independent of SIV infection, (3) the CD4 negative memory T cells maintain functions which are normally attributed to CD4 T cells including production of IL-2, production of IL-17, expression of FoxP3 and expression of CD40L (4) loss of CD4 expression protects these T cells from infection by SIV in vivo (5) these CD4 negative T cells maintain MHC-II restriction and (6) CD4 T cells do not need stimulation through the T cell receptor to down regulate CD4. Hence downregulation of CD4, with maintenance of CD4 T cell effector functions, is a likely mechanism by which AGM are able to live with SIVagm and avoid disease progression. We have also initiated studies to understand the molecular mechanism by which CD4 protein is down regulated as AGM nave CD4 T cells are stimulated to enter the memory pool. In collaboration with Dr. Remy Bosselut, we have begun investigating the expression pattern and genetic sequence of several transcription factors known to be critically important for CD4 and CD8 lineage selection. We have found significant alterations in the expression patterns of key transcription factors in AGM compared to mice (where these transcription factors are extensively studied). Moreover we have identified mutations in these transcription factors in AGM. MICROBIAL TRANSLOCATION STUDIES Progressive lentiviral infection of primates is associated with a multifaceted activation of the immune system. This chronic immune activation has been shown to be tightly associated with HIV and SIV disease progression. We recently showed that one of the major causes of this chronic immune activation was microbial translocation. During the acute phase of infection the structural and immunological barrier of the gastrointestinal tract are damaged and microbial products consequently translocate from the lumen into peripheral circulation. In circulation these microbial products can stimulate the adaptive and innate arms of the immune system. Indeed, we found significant correlations between microbial translocation and measures of immune activation in chronically HIV/SIV-infected individuals. To understand the mechanisms underlying microbial translocation we have investigated the integrity of the epithelial barrier and status of the GI tract immune system in SIV-infected Asian macaques. We have found that an inability to maintain a tight GI tract epithelial barrier is associated with alterations in the functionality and composition of lymphocytes within the lamina propria of the GI tract. We have also aggressively sequenced bacteria within the GI tracts of SIV-infected and uninfected animals and bacteria which are capable of translocating into tissues. We found that only specific bacteria are capable of translocating into tissues. Hence the identify of microbes within the GI tract is an important consideration. Therefore we have investigated adjuvant therapeutic interventions to ameliorate bacterial dysbiosis. We found significant improvement of the GI tract immune system after adjuvant treatment of antiretrovirals with probiotics in SIV-infected Asian macaques.